Characterization of the grass group I pollen allergens Cyn d 1from Cynodon dactylon
碩士 === 國立陽明大學 === 醫學生物技術研究所 === 90 === Many major allergens from mites and moulds were shown to be enzymes. However, few pollen allergens display enzymatic activities. There are controversial arguments whether the group I pollen allergen posses the protease activity. In this thesis, the g...
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ndltd-TW-090YM0006040202016-06-24T04:15:13Z http://ndltd.ncl.edu.tw/handle/84877682527072780046 Characterization of the grass group I pollen allergens Cyn d 1from Cynodon dactylon Cynd1蛋白性質及結構之初步分析 ChuanHsiang Huang 黃駿翔 碩士 國立陽明大學 醫學生物技術研究所 90 Many major allergens from mites and moulds were shown to be enzymes. However, few pollen allergens display enzymatic activities. There are controversial arguments whether the group I pollen allergen posses the protease activity. In this thesis, the group I allergen (Cyn d 1) from Cynodon dactylon was isolated and characterized. Cyn d 1 can be divided into acidic and basic groups. By using a monoclone antibody affinity column and from Pichia pastoris expression system both native and recombinant Cyn d 1 were isolated from Cynodon dactylon. Rapid degradation of isolated proteins implies that Cyn d 1 may contain protease activity. The substrate digestion experiment also reveals that Cyn d 1 can cleavage the C-terminal of arginine. Furthermore, the zymogram experiments showed that both basic and acidic Cyn d 1 could cleave Casein and Gelatin not BSA. Only EDTA can inhibit the protease activity when incubation with various inhibitors. Therefore, Cyn d 1 might be a metallo-proteinase. Structural prediction by 3D-PSSM suggested that the N-terminal domain of Cyn d 1 (144 amino acids) is similar to endoglucanase, and the C-terminal domain (100 amino acids) is similar to group 2 allergens. Both endoglucanase and group 2 allergens are β proteins. Consistent with the prediction, the CD measurement revealed β structure. Both N- and C-terminal domains were expressed in Pichia pastoris expression system and shown to contain protease activity by the zymogram experiments. ZoNan Chang ShwuHuey Liaw 張若南 廖淑惠 2002 學位論文 ; thesis 0 zh-TW |
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碩士 === 國立陽明大學 === 醫學生物技術研究所 === 90 === Many major allergens from mites and moulds were shown to be enzymes. However, few pollen allergens display enzymatic activities. There are controversial arguments whether the group I pollen allergen posses the protease activity. In this thesis, the group I allergen (Cyn d 1) from Cynodon dactylon was isolated and characterized.
Cyn d 1 can be divided into acidic and basic groups. By using a monoclone antibody affinity column and from Pichia pastoris expression system both native and recombinant Cyn d 1 were isolated from Cynodon dactylon. Rapid degradation of isolated proteins implies that Cyn d 1 may contain protease activity. The substrate digestion experiment also reveals that Cyn d 1 can cleavage the C-terminal of arginine. Furthermore, the zymogram experiments showed that both basic and acidic Cyn d 1 could cleave Casein and Gelatin not BSA. Only EDTA can inhibit the protease activity when incubation with various inhibitors. Therefore, Cyn d 1 might be a metallo-proteinase.
Structural prediction by 3D-PSSM suggested that the N-terminal domain of Cyn d 1 (144 amino acids) is similar to endoglucanase, and the C-terminal domain (100 amino acids) is similar to group 2 allergens. Both endoglucanase and group 2 allergens are β proteins. Consistent with the prediction, the CD measurement revealed β structure. Both N- and C-terminal domains were expressed in Pichia pastoris expression system and shown to contain protease activity by the zymogram experiments.
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ZoNan Chang |
author_facet |
ZoNan Chang ChuanHsiang Huang 黃駿翔 |
author |
ChuanHsiang Huang 黃駿翔 |
spellingShingle |
ChuanHsiang Huang 黃駿翔 Characterization of the grass group I pollen allergens Cyn d 1from Cynodon dactylon |
author_sort |
ChuanHsiang Huang |
title |
Characterization of the grass group I pollen allergens Cyn d 1from Cynodon dactylon |
title_short |
Characterization of the grass group I pollen allergens Cyn d 1from Cynodon dactylon |
title_full |
Characterization of the grass group I pollen allergens Cyn d 1from Cynodon dactylon |
title_fullStr |
Characterization of the grass group I pollen allergens Cyn d 1from Cynodon dactylon |
title_full_unstemmed |
Characterization of the grass group I pollen allergens Cyn d 1from Cynodon dactylon |
title_sort |
characterization of the grass group i pollen allergens cyn d 1from cynodon dactylon |
publishDate |
2002 |
url |
http://ndltd.ncl.edu.tw/handle/84877682527072780046 |
work_keys_str_mv |
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